乙型肝炎病毒大蛋白临床检测意义初探

目的通过检测乙型肝炎患者血清中乙型肝炎病毒大蛋白（Hepatitis B virus large surface protein,LHBs）、HBVDNA、乙肝五项（HBV M）等指标．探讨LHBs用于临床检测乙型肝炎的意义。方法采用酶联免疫吸附实验（ELISA）检测乙肝五项（HBVM）以及乙型肝炎病毒大蛋白（LHBs）,采用荧光定量PCR方法对标本HBV DNA进行检测。结果①LHBs吸光度（OD值）与HBV DNA呈正相关关系（γ=0．972）;②73例HBsAg、HBeAg均为阳性患者中HBV DNA与LHBs的检出没有显著性差异（χ^2=0．00,P〉0．05）。两者的检出一致率为97．26％[（71＋0）／73]。③80例HBsAg阳性、HBeAg阴性患者中HBV DNA与LHBs的检出没有显著性差异（χ^2=0．11,P〉0．05）。两者的检出一致率为82．19％[（42＋18）／73]。结论乙型肝炎病毒大蛋白（LHBs）与HBV DNA呈正相关性,临床上可以用于反映乙型肝炎病毒的复制情况,特别是HBeAg阴性的患者病毒复制情况,用于填补由于病毒变异引起的HBeAg检测的不足。     

慢性HBV感染患者血清cccDNA与HBVDNA、HBeAg关系研究

目的研究慢性HBV感染患者血清cccDNA与HBVDNA、HBeAg关系。方法采用分子信标PCR技术检测156例慢性HBV感染患者血清cccDNA。结果血清HBVDNA、HBeAg阳性组的血清cccDNA阳性率分别与阴性组相比较差异有显著性（P〈0．01）,血清HBVDNA阴性组血清cccDNA阳性率为0,与HBeAg阴性组比较差异也有显著性（P〈0．01）。HBeAg阳性组血清cccDNA水平较高,与阴性组比较差异有显著性（P〈0．01）。结论血清cccDNA与血清HBVDNA、HBeAg密切相关,是判断HBV病毒复制、活动、抗病毒效果的指标,可能比HBeAg更有价值。     

HBeAg阴性乙肝患者LHBs的检测及其意义

目的探讨检测乙型肝炎HBeAg阴性患者病毒表面大蛋白（LHBs）用于反映体内乙型肝炎病毒复制的意义。方法采用酶联免疫吸附实验（ELISA）和荧光定量PCR法对807例HBeAg阴性血清标本进行LHBs及HBVDNA检测。结果①在807例HBeAg阴性慢性乙肝血清中HBVDNA阳性率为54．40％（439／807），LHBs的阳性率为56．75％（458／807），二者比较无显著性差异（Χ^2=0．813，P〉0．05），两者总体符合率为91．70％（740／807）；②HBeAg阴性慢性乙肝不同HBV—M模式的HBV—DNA与LHBs检出结果均无显著性差异（P〉0．05）。⑨LHBs含量与HBVDNA拷贝数呈正相关（r=0．995，P〈0．001）。结论LHBs与HBVDNA具有良好的相关关系，能够反映出HBeAg阴性乙肝患者体内HBV复制的程度，是HBVDNA复制水平判定的有效指标，具有重要的临床意义。     

HBeAg阴性HBV DNA阳性HBV感染者血清HBV DNA与ALT关系之探讨

目的探讨乙型肝炎病毒e抗原（HBeAg）阴性乙型肝炎病毒（HBV）DNA阳性HBV感染者血清HBVDNA与丙氨酸转氨酶（ALT）之间的关系。方法对508例HBeAg阴性HBVDNA阳性（10^3～10^8copies／mL）HBV感染者血清进行ALT测定。结果乙型肝炎“小三阳”（HBsAg＋／HBeAb＋／HBcAb＋）患者血清ALT增高率：10^3～10^4copies／mL组为56．7％，10^5～10^6copies／mL组为80．5％，10^7～10^8copies／mL组为94．7％;HBsAg＋／抗-HBc＋患者血清ALT增高率：10^3～10^4 copies／mL组为64．3％，10^5～10^6copies／mL组为83．0％，10^7～10^8 copies／mL组为100．0％。结论HBeAg阴性HBVDNA阳性HBV感染者血清HBVDNA与ALT之间有显著相关性，因此HBVDNA可以作为HBeAg阴性HBV感染者肝细胞损害程度的评估指标。     

前S1抗原在乙型肝炎诊断中的应用研究

目的研究在不同病情乙肝患者血清中前S1抗原（HBVpre-S1）水平的变化与HBV血清标志物、HBV-DNA的临床相关性,评价HBVpre—S1的检测在乙型肝炎诊断中的应用价值。方法用酶联免疫吸附法（ELISA）分别对268例HBV血清进行五项血清标志物,Pre—S1的检测。用全自动生化分析仪赖氏法检测肝功能,同时采用PCR荧光定量法检测HBV-DNA的量。结果196例慢性乙肝（CH）患者中,HBeAg（＋）组和HBeAg（-）组比较,Pre-S1（＋）和HBVDNA（＋）检出率及Pre-S1／HBsAg的相对比值均高于HBeAg（-）组,差异具有统计学意义P〈0．01;且在HBeAg（-）组中。HBeAb（-）组和HBeAb（＋）组相比,Pre-S1／HBsAg相对比值也显著高于HBeAb（＋）组,P〈0．05;在CH患者中,轻、中、重三组间的Pre-S1（＋）率,HBVDNA及Pre-S1／HBsAg相对比值相比较,差异均无统计学意义,且Pre-S1／HBsAg相对比值水平和肝脏炎症程度无关。在72例AH患者中,Pre-S1／HBsAg的相对比值与肝脏急性损伤的严重程度相关,在AH患者Pre-S1／HBsAg相对比值水平下降越早越快,病程越短,预后越好。结论Pre-S1可作为HBV复制的标志物;而Pre-S1／HBsAg的相对比值的高低,在肝炎的诊断,AH预后的判断方面具有重要的价值;联合检测Pre-S1可补充和完善HBV五项检测的不足,尤其对HBeAg（-）或者变异的HBV感染者能更好地反映病毒的复制状态和传染性。     

乙型肝炎病毒前S1抗原与HBV DNA及肝功能的关系研究

目的探讨不同模式乙肝患者血清中乙肝病毒前S1抗原（HBVPreS1—Ag）与HBVDNA及ALT、AST的相互关系，分析HBVPreS1—Ag用于乙肝临床诊断的意义。方法采用酶联免疫吸附实验（ELISA）检测350例乙肝患者血清HBVPreS1—Ag，荧光定量PCR检测HBVDNA，全自动生化分析仪检测ALT和AST。结果相同乙肝模式患者血清HBVPreS1-Ag与HBVDNA检出率的差异无显著性；HBVPreS1-Ag与HBVDNA、HBeAg与HBVPreS1-Ag、HBeAg与HBVDNA的符合率分别为86．9％（K=0．72，P〈0．05）、75．1％（K=0．59，P〈0．05）、79．7％（K=0．62，P〈0．05）；HBVPreS1-Ag阳性患者血清ALT和AST升高率和水平均高于HBVPreS1-Ag阴性患者。结论HBVPreS1—Ag与HBV—DNA有较高的符合率，可作为反映HBV复制的指标。而且，HBVPreS1—Ag阳性患者血清ALT和AST水平高于阴性患者，表明HBVPreS1—Ag可反映肝功能状况。     

检测乙肝表面抗原大蛋白的临床意义

[目的]检测乙肝患者血清中乙肝表面抗原大蛋白（ Hepatitis B Virus Large Surface Protein, LHBs） 、乙肝前SI、HBVDNA以及ALT，探讨LHBs用于乙肝患者临床诊断的意义。[方法]采用酶联免疫吸附实验（ELISA）检测LHBs以及乙肝前st，采用化学发光方法检测乙肝两对半，采用荧光定量PCR方法对患者HBVDNA进行检测，采用全自动生化分析仪检测丙氨酸转氨酶（ALT）。[结果]1．相同乙肝模式患者血清LHBs与HBVDNA检出率无显著差异（P〉0．05）；2．HBeAg阳性患者血清中LHBs与HBVDNA阳性率均明显高于HBeAg阴性患者，差异显著（P〈0．05）；3．相同乙肝模式患者血清乙肝前S1的检出低于HBVDNA的检出，两者差异显著（P〈0．05）；4．LHBs阳性患者的ALT明显高于LHBs阴性患者。[结论]乙肝表面抗原大蛋白（LHBs）可以弥补由于乙肝病毒变异引起的HBeAg检测的不足，并在一定程度上反映了乙肝患者肝功能状况。     

乙型肝炎病毒共价闭合环状DNA检测方法的建立及应用

[目的]建立和应用半巢式聚合酶链反应法（PCR）检测慢性乙型肝炎患者血清中乙型肝炎病毒（HBV）共价闭合环状DNA（cccDNA）。[方法]根据HBV基因组DNA正负链上下游缺口序列，设计3个相对保守的引物，建立HBV cccDNA半套式PCR，并用该法检测96例慢性乙型肝炎患者血清中HBV cccDNA。[结果]96例慢性乙型肝炎患者中，35例HBV cccDNA阳性，阳性率为36．5％。HBV cccDNA阳性组的HBeAg、HBV DNA、ALT和AST水平均明显高于阴性组（P〈0．05），但两组的HBsAg水平无显著差异（P〉0．05）。HBV DNA≥10^5组的HBV cccDNA检出率为58．3％（35／60），明显高于HBVDNA〈10^5组（0％，0／36）（P=0．000）；HBeAg阳性组的HBV cccDNA检出率为50．9％（28／55），明显高于HBeAg阴性组（7／41，17．1％）（P=0．001）。[结论]半套式PCR方法简便，可用于测定血清中HBV cccDNA，以评价抗病毒治疗慢性乙型肝炎的疗效。     

乙型肝炎病人及其配偶HBV血清标志物检测

目的比较分析乙型肝炎病人及其配偶HBV血清标志物的检测结果。方法采用ELISA法和荧光PCR法对316例乙型肝炎病人及其配偶进行乙型肝炎五项指标与HBVDNA定量检测。结果HBVDNA的阳性率和含量在HBeAg（＋）与HBeAg（-）组差异均有显著意义（χ^2=31．97、68．79,P〈0．01）;HBsAg的相对滴度与HBVDNA的含量关系不密切。乙型肝炎病人配偶HBV感染率为58．2％,有4例HBVDNA含量〉1．0×10^6 copies／L。结论HBeAg与HBV的复制和HBVDNA的高含量关系密切,而HBsAg的相对滴度则不能反映HBV的复制程度及传染性强弱。配偶间的密切接触可以引起HBV感染,但很少导致HBVDNA在体内的存在和复制。     

HBV—DNA检出情况与乙肝五项结果之间的相关性研究

[目的]探讨HBV—DNA的检测结果与乙肝两对半结果之间的相关性。[方法]运用荧光定量PCR法检测225例疑似乙肝患者血清中的HBV—DNA含量,同时采用ELISA法检测乙肝两对半标志物,并对结果进行相关性分析。[结果]HBsAg（＋）HBeAg（＋）HBcAb（＋）组（大三阳组）患者血清中HBVDNA的阳性率为85％（51／60）,平均拷贝数为2．24×10^7 copies／mt;HBsAg（＋）HBeAb（＋）HBcAb（＋）组（小三阳组）患者血清中HBVDNA的阳性率为31％（31／98）,平均拷贝数为3．86×10^5copies／ml;HBsAg（＋）HBcAb（＋）组患者血清中HBVDNA的阳性率为38％（5／13）,平均拷贝数为6．93×10^5copies／ml。[结论]大三阳组和小三阳组阳性率相比差异有统计学意义（x^2=42．458,P〈0．005）;大三阳组和HBsAg（＋）HBcAb（＋）组阳性率相比差异有统计学意义（x^2=12．954,P〈0．005）;HBVDNA病毒含量检测更能准确反映患者HBV感染情况,因此在检测中应该联合使用两种方法,提高检测的准确性,指导临床对患者的病情进行诊断、指导治疗和观察疗效。     

战斗机飞行员血清胆红素与血清铁、血红蛋白等指标的相关性研究

目的探讨战斗机飞行员血清胆红素、血清铁和血红蛋白含量等指标变化的机制及相关性。方法所选人员分为战斗机飞行员（观察组）83名与同期入院地面干部（对照组）30名,测定两组的血清胆红素、血清铁、血红蛋白等指标并进行比较和相关性分析;战斗机飞行员又分为高性能战斗机飞行员组47名和歼击机飞行员组36名,比较其血清胆红素、血清铁、血红蛋白等指标的变化。结果①战斗机飞行员血清胆红素、血清铁、血红蛋白均降低（P〈0.01或P〈0.05）,其中高性能战斗机飞行员更加明显。②血清胆红素与血红蛋白呈正相关（P〈0.01）,与血清铁无相关性（P〉0.05）;血红蛋白与血清铁呈正相关（P〈0.01）。结论战斗机飞行员血清胆红素水平降低,可促进动脉粥样硬化形成,增加冠心病发生的危险性;由于血清铁降低,存在铁缺乏,致使血红蛋白含量下降,导致缺氧耐受力降低。因此,及早采取干预措施,对于降低飞行员冠心病发生的危险性及延长飞行寿命有重要意义。     

Accuracy of serum ferritin determinations in tissue preparations and human serum

Introduction of the NBSB 80/602 reference preparation for the calibration of ferritin immunoassays has reduced the inter-assay variability, and represents the first important step towards standardisation of ferritin immunoassays. However, our investigations show that comparison between assay results is still impossible, owing to differences in assay methodology and performance, differences in the specificities of the antibodies and antigens used and other possible interfering substances. We conclude that all kits detect mainly the more basic isoferritins in serum, and that all isoferritins (acidic, intermediate and basic) are systematically underestimated. Since we also showed possible immunological differences between reference or kit standards and serum ferritin, we conclude that the present kits have poor accuracy. To diminish inter-assay variability and to increase the accuracy of serum ferritin determinations, a method is needed that detects all basic, intermediate and acidic isoferritins and measures the true ferritin concentration in serum under normal and pathological circumstances. This reference method can be used to evaluate interference and systematic errors in routine methods. The introduction of a reference method, in combination with the NBSB 80/602 human liver reference preparation, is the second important step towards the accurate standardisation of ferritin immunoassays.     

Radioimmunological determination of reverse triiodothyronine in unextracted serum and serum dialysates

A sensitive radioimmunoassay for measurements of 3,3'-5'-triiodothyronine (reverse T3, rT3) in small amounts of unextracted serum is described. The interference of rT3 binding proteins in serum was precluded by addition of 8-anilino-1-naphthalene sulphonic acid (ANS). The cross reaction of T4 with the rT3 anti-serum varied with the concentration of T4 in the samples. At 50 per cent inhibition of [125I]rT3 binding, the T4 cross reaction was 0.075% (mol/mol). All values were corrected for T4 cross reaction. By the present method total rT3 averaged 0.37 nmol/l in thirty-two normal subjects. Higher values (0.81-1.98 nmol/l) were obtained in seventeen thyrotoxic patients, while the rT3 was very low (less than or equal to 0.046 nmol/l) in ten patients with severe primary hypothyroidism. A modification of the total rT3 assay was used for measurements of rT3 in serum dialysates (free rT3). The sensitivity was 0.46 pmol/l. This sensitivity did not allow detection of free rT3 in all normal subjects.     

血清胆红素水平与冠状动脉狭窄病变程度及血脂的关系

目的 探讨血清胆红素水平与冠状动脉狭窄病变程度及血脂的关系。方法 将152例已行冠状动脉造影患者分为冠心病组（狭窄≥50％）及对照组（冠状动脉造影正常者）,比较两组间血清总胆红素（TBIL）、直接胆红素（DBIL）、间接胆红素（IBIL）及血脂水平,并对胆红素与血脂进行直线相关分析。结果冠心痛组血清TBIL低于对照组（P＜0．05）,总胆固醇（TC）、甘油三酯（TG）和低密度脂蛋白胆固醇（LDL—C）均高于对照组（P＜0．05或P＜0．01。冠心病组中单支病变和中度狭窄患者血清TBIL和DBIL水平均低于对照组（P＜0．05）,多支病变和重度狭窄患者血清TBIL、DBIL和IBIL水平低于单支病变和中度狭窄患者以及对照组（P＜0．05或P＜0．01）。相关分析显示,血清胆红素与Tc、TG和LDL—C呈显著负相关。结论 胆红素影响血脂代谢。低胆红素血症患者易患冠心病,血清胆红素水平越低．冠状动脉狭窄病变程度越重．     

血清肝纤维化标志物与肝功能指标的相关性研究

目的 研究肝硬化患者肝纤维化血清标志物与肝功能指标的关系.方法 62例肝硬化患者检测透明质酸(HA)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(ⅣC)、层粘连蛋白(LN)及肝功能指标,并进行Child Pugh分级和分析.结果 经相关分析和秩和检验,肝纤维化指标与胆碱酯酶(CHE)、凝血酶原活动度(PTA)呈负相关(P＜0.05～0.01),与胆红素(TBIL)、谷草转氨酶(AST)呈正相关(P＜0.05～0.01),并随Child Pugh评分增加而递增(P＜0.05～P＜0.01).结论 肝纤维化检测与肝功能指标明确相关,综合多项无创性检查诊断肝纤维化是临床的需要.     

Comparison of the relationships between serum apolipoprotein B and serum lipid distributions

BACKGROUND: Apolipoprotein B (apo B) has been reported to be a better predictor of coronary artery disease than cholesterol indices. The objectives of this study were to evaluate concordances/discordances between cholesterol indices and apo B and to assess the factors that influence them. METHODS: For this study, 11 816 individuals (6965 males, 4851 females), none of whom had a past history of coronary artery disease, were selected from among visitors to the health promotion center at Kangbuk Samsung Hospital between January and December 2002. We assessed concordances between the biochemical indices of atherogenicity and evaluated factors associated with discordances. RESULTS: Apo B and various cholesterol indices were correlated, although concordance fell within the range 47%-56%. Multinomial logistic regression analysis showed an increasing risk of a disproportionately higher apo B than LDL-cholesterol in males, the elderly, smokers, individuals with metabolic syndrome, in those with high HDL-cholesterol or triglyceride (TG) concentrations or larger waist circumferences, and in those with low total cholesterol (TC). CONCLUSIONS: The introduction of apo B to standard lipid profile testing could improve the evaluation of risk factors of coronary artery disease and aid more accurate assessment of the effects of cholesterol-lowering therapy, particularly in males, the elderly, smokers, or in individuals with metabolic syndrome, high HDL-cholesterol, high TGs, larger waist circumferences, or low TC.     

The influence of serum and serum proteins on calcium oxalate crystal growth and aggregation

The effects of serum, albumin, alpha-globulin, and a mixture of alpha- and beta-globulin on the growth and aggregation of calcium oxalate crystals were measured in a standard seeded crystallisation system. At concentrations below 0.01% blood, which corresponded to microscopic haematuria, all were potent inhibitors of crystal aggregation whilst only having a minor effect on crystal growth. It was found that albumin, alpha-globulin and beta-globulin can account for the total inhibitory effect of serum on crystal growth and crystal aggregation.     

Concentrations and levels of activity of important serum protein components in preserved serum

The content of transport proteins, immune globulins, components of the complement system and protease inhibitors were estimated in two different plasma preparations, using bioassays by chromogenic substrates and laser nephelometry. The content of proteins under consideration was found about 50% less than in untreated human plasma. Some proteins such as antithrombin III and factor XIII are missing. The results are discussed in respect to clinical used of plasma preparations.